Non-viral Induction of Transgene-free iPSCs from Somatic Fibroblasts of Multiple Mammalian Species [pig]

Induced pluripotent stem cells (iPSCs) are capable of providing an unlimited source of cells from all three germ layers as well as germ cells. The derivation and usage of iPSCs from various animal models may facilitate stem-cell-based therapy, generation of gene-modified animals, and evolutionary studies assessing interspecies differences. However, there is a lack of species-wide methods for deriving iPSCs, in particular by means of non-viral and non-transgene-integrating (NTI) approaches. Here, we demonstrated the derivation of iPSCs from somatic fibroblasts of multiple mammalian species from three different taxonomic orders, including the common marmoset (Callithrix jacchus) in Primates, the dog (Canis lupus familiaris) in Carnivora, and the pig (Sus scrofa) in Cetartiodactyla, by combinatorial usage of chemical compounds and NTI episomal vectors. Interestingly, the somatic fibroblasts temporarily acquired a neural stem cell (NSC)-like state during the reprogramming procedure. Collectively, our method, robustly applicable to various species, holds a great potential for facilitating stem-cell-based research using various animals in Mammalia. Overall design: RNA seq of porcine fibroblasts, iNSLCs, and iPSCs

诱导多能干细胞（Induced pluripotent stem cells，iPSCs）可提供源自三胚层及生殖细胞的无限细胞来源。从多种动物模型中获取并应用iPSCs，有助于推动干细胞治疗、基因修饰动物构建以及评估物种间差异的进化生物学研究。然而，当前仍缺乏适用于全物种的iPSCs制备方法，尤其是非病毒非转基因整合（non-transgene-integrating，NTI）技术手段。本研究通过化学小分子与NTI附加型载体的联合使用，成功从三个不同分类目下的多种哺乳动物体细胞成纤维细胞中诱导获得iPSCs，涉及灵长目普通狨猴（Callithrix jacchus）、食肉目家犬（Canis lupus familiaris）以及鲸偶蹄目家猪（Sus scrofa）。有趣的是，在重编程过程中，体细胞成纤维细胞会短暂呈现神经干细胞（neural stem cell，NSC）样状态。综上，本研究所建立的方法可稳定适配多种物种，具备推动哺乳类动物干细胞相关研究的巨大应用潜力。实验设计：对家猪成纤维细胞、诱导神经干细胞样细胞（iNSLCs）及iPSCs进行RNA测序。